Procedure for crystallization of (22E)-(24R)-2-methylene-22-dehydro-1α,24-25-trihydroxy-19-nor-vitamin D3

Abstract

Disclosed are methods of purifying the compound (22E)-(24R)-2-methylene-22-dehydro-1α,24,25-trihydroxy-19-nor-vitamin D 3 to obtain the compound in crystalline form. The methods typically include the steps of dissolving (22E)-(24R)-2-methylene-22-dehydro-1α,24,25-trihydroxy-19-nor-vitamin D 3 in a solvent comprising ethyl acetate and hexane to form a solution, allowing crystals of (22E)-(24R)-2-methylene-22-dehydro-1α,24,25-trihydroxy-19-nor-vitamin D 3 to form and precipitate from the solution, and recovering the crystals of (22E)-(24R)-2-methylene-22-dehydro-1α,24,25-trihydroxy-19-nor-vitamin D 3 from the solution.

We claim: 
     
       1. A compound having the formula 
       
         
           
           
               
               
           
         
       
       in a crystalline form and named (22E)-(24R)-2-methylene-22-dehydro-1α,24,25-trihydroxy-19-nor-vitamin D 3 , wherein the crystalline form has molecular packing arrangement defined by space group P2 1  and unit cell dimensions: a=16.3058(9) Å; b=10.0461(6) Å; c=26.0527(15) Å; α=90°; β=109.001(3)°; and γ=90°. 
     
     
       2. A three dimensional structure for (22E)-(24R)-2-methylene-22-dehydro-1α,24,25-trihydroxy-19-nor-vitamin D 3  as defined by the molecular packing arrangement set forth in  claim 1 . 
     
     
       3. A method of purifying (22E)-(24R)-2-methylene-22-dehydro-1α,24,25-trihydroxy-19-nor-vitamin D 3 , comprising the steps of:
 (a) dissolving (22E)-(24R)-2-methylene-22-dehydro-1α,24,25-trihydroxy-19-nor-vitamin D 3  in a volume of ethyl acetate as solvent to prepare a solution of (22E)-(24R)-2-methylene-22-dehydro-1α,24,25-trihydroxy-19-nor-vitamin D 3 , 
 (b) adding a volume of hexane dropwise to the solution, to reach a saturation point, to form a solution of (22E)-(24R)-2-methylene-22-dehydro -1α,24,25-trihydroxy-19-nor-vitamin D 3  in a mixture of ethyl acetate and hexane as solvent wherein crystals of (22E)-(24R)-2-methylene-22-dehydro-1α,24,25-trihydroxy-19-nor-vitamin D 3  form and precipitate from the solution; and 
 (c) separating the (22E)-(24R)-2-methylene-22-dehydro-1α,24,25-trihydroxy-19-nor-vitamin D 3  crystals from the solution, wherein the (22E)-(24R)-2-methylene-22-dehydro-1α,24,25-trihydroxy-19-nor-vitamin D 3  crystals have a crystalline form that has molecular packing arrangement defined by space group P2 1  and unit cell dimensions: a=16.3058( 9 ) Å; b=10.0461(6) Å; c=26.0527(15) Å; α=90°; β=109.001(3)°; and γ=90°. 
 
     
     
       4. The method of  claim 3 , wherein the solution of (22E)-(24R)-2-methylene-22-dehydro-α,24,25-trihydroxy-19-nor-vitamin D 3  in the mixture of ethyl acetate and hexane as solvent is placed at room temperature, wherein crystals of (22E)-(24R)-2-methylene-22-dehydro-1α,24,25-trihydroxy-19-nor-vitamin D 3  form and precipitate from the solution. 
     
     
       5. The method of  claim 3 , wherein the step of separating comprises filtering the mixture and precipitate to obtain the crystals. 
     
     
       6. The method of  claim 3  including a further step (d) comprising repeating steps (a) through (c) using the recovered crystals of (22E)-(24R)-2-methylene-22-dehydro-1α,24,25-trihydroxy-19-nor-vitamin D 3  from step (c) in step (a). 
     
     
       7. The method of  claim 3  wherein the mixture of ethyl acetate and hexane comprises about 20-30% ethyl acetate and about 70-80% hexane by volume. 
     
     
       8. A pharmaceutical composition containing an effective amount of the compound of  claim 1  in crystalline form and a pharmaceutically acceptable excipient. 
     
     
       9. A method of treating or preventing a skin disease, disorder, or condition in a subject in need thereof, the method comprising administering to the subject an effective amount of the compound of  claim 1  in crystalline form. 
     
     
       10. The method of  claim 9 , wherein the skin disease, disorder, or condition is psoriasis. 
     
     
       11. A method of treating or preventing a cell proliferative disease or disorder in a subject in need thereof, the method comprising administering to the subject an effective amount of the compound of  claim 1  in crystalline form. 
     
     
       12. The method of  claim 11 , wherein the cell proliferative disease or disorder is selected from the group consisting of leukemia, neuroblastoma, retinoblastoma, melanoma, colon cancer, breast cancer, and prostate cancer. 
     
     
       13. A method of treating or preventing a disease or disorder in a subject in need thereof, the disease or disorder selected from the group consisting of multiple sclerosis, diabetes mellitus, lupus, host versus graft reaction, and rejection of transplants, the method comprising administering to the subject an effective amount of the compound of  claim 1  in crystalline form. 
     
     
       14. A method of treating or preventing an inflammatory disease or disorder in a subject in need thereof, the method comprising administering to the subject an effective amount of the compound of  claim 1  in crystalline form. 
     
     
       15. The method of  claim 14 , wherein the inflammatory disease or disorder is selected from the group consisting of rheumatoid arthritis, asthmas, and inflammatory bowel diseases. 
     
     
       16. The method of  claim 14 , wherein the inflammatory disease or disorder is selected from the group consisting of Crohn's disease and ulcerative colitis. 
     
     
       17. A method of treating or preventing obesity, inhibiting adipocyte differentiation, inhibiting SCD-1 gene transcription, and/or reducing body fat in a subject in need thereof, the method comprising administering to the subject an effective amount of the compound of  claim 1  in crystalline form.